Surface enhanced raman scattering substrate assembly

ABSTRACT

The present disclosure provides a surface enhanced Raman scattering substrate assembly for detecting an analyte. The assembly can include an etched fiber base. The assembly can further include a metallic nanoparticle coating disposed over at least a portion of the surface etched fiber base.

CROSS REFERENCE TO RELATED APPLICATION

This application is a continuation of U.S. patent application Ser. No.16/439,186, filed Jun. 12, 2019, which claims the benefit of priority toU.S. Patent Provisional Application No. 62/685,102, filed Jun. 14, 2018,which are incorporated by reference herein in their entireties.

STATEMENT OF GOVERNMENT SUPPORT

This invention was made with Government support under Grant #USDA-NIFA2016-67017-24458 awarded by the National Institute of Food andAgriculture of the United States Department of Agriculture. The U.S.Government has certain rights in this invention.

BACKGROUND

Surface enhanced Raman scattering (SERS) can be useful for manydifferent applications. For example, SERS can be used to detect a widevariety of biomolecules, metabolites, or other materials. However, alack of portability of SERS devices and the ability to accomplishreal-time detection of analytes are problems with some SERS systems.There is a need therefore, for improving the portability and real-timedetection of SERS systems.

SUMMARY OF THE DISCLOSURE

The present disclosure provides a surface enhanced Raman scatteringsubstrate assembly for detecting an analyte. The assembly can include anetched fiber base. The assembly can further include a metallicnanoparticle coating disposed over at least a portion of the surfaceetched fiber base.

The present disclosure further provides a method for detecting ananalyte. The method includes contacting an etched fiber base with amedium. The method further includes contacting the medium with anelectromagnetic emission. The method further includes detecting theanalyte and generating a spectrum.

The present disclosure further provides a method of making a surfaceenhanced Raman scattering substrate assembly for detecting an analyte.The assembly can include an etched fiber base. The assembly can furtherinclude a metallic nanoparticle coating disposed over at least a portionof the surface etched fiber base. The method includes etching a fiber toform an etched fiber base. The method further includes coating metallicnanoparticles on the surface of the etched fiber base.

BRIEF DESCRIPTION OF THE FIGURES

The drawings illustrate generally, by way of example, but not by way oflimitation, various embodiments discussed in the present document.

FIG. 1 is a schematic depiction of a surface enhanced Raman scatteringsubstrate assembly, in accordance with various embodiments.

FIGS. 2A-2B are a schematic depictions of a needle for a SERS substrateassembly, in accordance with various embodiments.

FIG. 3 is a schematic depiction of a method of making and using asurface enhanced Raman scattering substrate assembly, in accordance withvarious embodiments.

FIGS. 4A-4H are scanning electron microscope (SEM) images showing etchedfibers and coated etched fibers, in accordance with various embodiments.

FIG. 5 shows spectra generated from the surface enhanced Ramanscattering substrate assembly, in accordance with various embodiments.

FIGS. 6A-6B are a spectrum and concentration chart generated from thesurface enhanced Raman scattering substrate assembly, in accordance withvarious embodiments.

FIGS. 7A-7B are spectra generated from the surface enhanced Ramanscattering substrate assembly, in accordance with various embodiments.

FIG. 8 is a surface enhanced Raman scattering substrate assembly, inaccordance with various embodiments.

FIG. 9 are spectra generated from the surface enhanced Raman scatteringsubstrate assembly, in accordance with various embodiments.

FIG. 10 is a schematic depiction of the surface enhanced Ramanscattering substrate assembly located partially within a tomato, inaccordance with various embodiments.

FIG. 11 is a spectrum generated from the surface enhanced Ramanscattering substrate assembly deployed in the tomato of FIG. 10, inaccordance with various embodiments.

FIG. 12 is an SEM image showing pore size of an in-situ filter, inaccordance with various embodiments.

DETAILED DESCRIPTION

Reference will now be made in detail to certain embodiments of thedisclosed subject matter, examples of which are illustrated in part inthe accompanying drawings. While the disclosed subject matter will bedescribed in conjunction with the enumerated claims, it will beunderstood that the exemplified subject matter is not intended to limitthe claims to the disclosed subject matter.

Throughout this document, values expressed in a range format should beinterpreted in a flexible manner to include not only the numericalvalues explicitly recited as the limits of the range, but also toinclude all the individual numerical values or sub-ranges encompassedwithin that range as if each numerical value and sub-range is explicitlyrecited. For example, a range of “about 0.1% to about 5%” or “about 0.1%to 5%” should be interpreted to include not just about 0.1% to about 5%,but also the individual values (e.g., 1%, 2%, 3%, and 4%) and thesub-ranges (e.g., 0.1% to 0.5%, 1.1% to 2.2%, 3.3% to 4.4%) within theindicated range. The statement “about X to Y” has the same meaning as“about X to about Y,” unless indicated otherwise. Likewise, thestatement “about X, Y, or about Z” has the same meaning as “about X,about Y, or about Z,” unless indicated otherwise.

In this document, the terms “a,” “an,” or “the” are used to include oneor more than one unless the context clearly dictates otherwise. The term“or” is used to refer to a nonexclusive “or” unless otherwise indicated.The statement “at least one of A and B” has the same meaning as “A, B,or A and B.” In addition, it is to be understood that the phraseology orterminology employed herein, and not otherwise defined, is for thepurpose of description only and not of limitation. Any use of sectionheadings is intended to aid reading of the document and is not to beinterpreted as limiting; information that is relevant to a sectionheading may occur within or outside of that particular section.

In the methods described herein, the acts can be carried out in anyorder without departing from the principles of the disclosure, exceptwhen a temporal or operational sequence is explicitly recited.Furthermore, specified acts can be carried out concurrently unlessexplicit claim language recites that they be carried out separately. Forexample, a claimed act of doing X and a claimed act of doing Y can beconducted simultaneously within a single operation, and the resultingprocess will fall within the literal scope of the claimed process.

The term “about” as used herein can allow for a degree of variability ina value or range, for example, within 10%, within 5%, or within 1% of astated value or of a stated limit of a range and includes the exactstated value or range.

The term “substantially” as used herein refers to a majority of, ormostly, as in at least about 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%,98%, 99%, 99.5%, 99.9%, 99.99%, or at least about 99.999% or more, or100%.

Analysis of target compounds (analytes) from complex matrices, such asfood or biological samples, is challenging with traditional analyticalmethods. In particular, interference from other components in thematrices can occur during analysis. For this reason, many analyticalprocesses involved complicated and multi-step sample preparations toimprove sensitivity of the analysis. These processes can involveinvasive and destructive sampling.

Discussed herein is a method and assembly, in various embodiments, thatallows for high speed analysis of analytes in complex matrices withoutdestructive or invasive sampling. The method and assembly can include,in various embodiments, a micro-extraction device enabling in-situextraction and detection of analytes using surface enhanced Ramanspectroscopy (SERS) technology.

FIG. 1 is a schematic depiction of SERS substrate assembly 100. Surfaceenhanced Raman scattering substrate assembly 100 can include manysuitable components for detecting an analyte. Examples of suchcomponents for surface enhanced Raman scattering substrate assembly 100include etched fiber base 102 and metallic nanoparticle coating 104,which is disposed over at least a portion of the surface of etched fiberbase 102. Needle 106 at least partially circumscribes etched fiber base102. Needle 106 is an optional component. If needle 106 is present,needle 106 may be any suitable needle such as a hypodermic needlecapable of puncturing a material. Optionally, needle 106 can be used topuncture an opening in the medium, then removed before etched fiber base102 is inserted. In some embodiments, assembly 100 is in a transparentcontainer to allow SERS analysis.

Etched fiber base 102 can include any suitable material. Factors toconsider in choosing the material include the ability etch the material,the durability of the material at elevated temperatures, and the abilityof the material to participate in a reduction reaction during coating ofthe nanoparticles of metallic nanoparticle coating 104. An example of asuitable material for etched fiber base 102 includes stainless steel. Inother embodiments, etched fiber base 102 may include copper, lead,chromium, tin, magnesium, aluminum, zinc, manganese, calcium, alloysthereof, and mixtures thereof.

Etched fiber base 102 can have any elongated suitable shape. Forexample, etched fiber base 102 can be substantially cylindricallyshaped, substantially conically shaped, or substantially rectangularshaped. Etched fiber base 102 can have any suitable dimensions withrespect to length Li or width Wi. The term “length” is meant to apply toa largest dimension of etched fiber base 102. In some embodiments theetched fiber base has a length ranging from about 3 cm to about 6 cm.The term “width” applies to a largest dimension of etched fiber base 102substantially orthogonal to the length. In embodiments where etchedfiber base 102 is substantially cylindrically shaped, the width maycorrespond to a largest diameter of etched fiber base 102. In someembodiments the etched fiber base has a width ranging from about 100 μmto about 400 μm or from about 0.5 cm to about 5 cm. Etched fiber base102 can be, for example, from about 1 cm to about 10 cm (e.g., about 3cm to about 6 cm). Etched fiber base 102 can have, for example, athickness of about 50 μm to about 400 μm (e.g., about 50 μm to about 100μm). Fiber length and thickness can alternatively be determined based onthe sample vial or the sample itself, in addition to the needle size ifa needle is used.

Etched fiber base 102 can be etched to form a predetermined pattern or arandom pattern of grooves, depressions, ridges, pores, or the like.Etching can be accomplished using any suitable method such as acidetching or laser etching. Etching patterns may be formed using a screenor mask to selectively expose certain regions to the etchant.

Etching can be desirable for several reasons. For example, etching afiber increases the surface area of the fiber as compared to acorresponding fiber that is free of etching, or etched to a lesserdegree, that shares substantially the same length and width as etchedfiber base 102. The increased surface area can be helpful in embodimentswhere surface enhanced Raman scattering substrate assembly 100 is usedto detect an analyte in a liquid or gaseous phase. This can be becausethe increased surface area increases the contact points that the analytehas available to interact with upon metallic nanoparticle coating 104.The increased surface area on etched fiber base 102 can further allowfor an increased number of metallic nanoparticles to be included inmetallic nanoparticle coating 104.

Metallic nanoparticle coating 104 is dispersed over about 50% to about100% of the total surface area of etched fiber base 102, about 70% toabout 100%, about 90% to about 98%, or less than, equal to, or greaterthan about 70%, 75, 80, 85, 90, 95, or about 100%. Metallic nanoparticlecoating 104 includes a plurality of metallic nanoparticles. The totalsurface area of etched fiber base 102 is the total external surface areainclusive of pores or other non-planar aspects of the surface.

Individual metallic nanoparticles can include any suitable material. Forexample, individual metallic nanoparticles of the plurality ofnanoparticles can include Ag₂O, elemental silver, elemental gold,elemental copper, elemental platinum, mixtures thereof, alloys thereof,or combinations thereof. In some embodiments, each metallic nanoparticleis the same material. For example, in some embodiments of surfaceenhanced Raman scattering substrate assembly 100 each metallicnanoparticle can include elemental gold.

Individual metallic nanoparticles can have any suitable morphology. Forexample, individual metallic nanoparticles can have a morphology such asa nanosphere, a nanochain, a nanoreef, a nanobox, or a nanostar. In someembodiments, each metallic nanoparticle of metallic nanoparticle coating104 can have the same morphology. In other embodiments, however,metallic nanoparticle coating 104 can include a mixture of metallicnanoparticles having different morphologies. In some embodiments,surface enhanced Raman scattering substrate assembly 100 can furtherinclude a layer of metallic microparticles disposed on etched fiber base102. Microparticles are generally understood to refer to particlesindividually having at least one dimeson (e.g., length, width,thickness, diameter, or height) in the micrometer range. The micrometerrange can include any distance from about 1 μm to about 1000 μm, about100 μm to about 500 μm, or less than, equal to, or greater than about 1μm, 100, 200, 300, 400, 500, 600, 700, 800, 900, or about 1000 μm.Nanoparticles are generally understood to refer to individual particleshaving at least one dimension (e.g., length, width, thickness, diameter,or height) in the nanometer range. The nanometer range can include anydistance from about 1 nm to about 10000 nm, about 100 nm to about 500nm, or less than, equal to, or greater than about 1 nm, 1000, 2000,3000, 4000, 5000, 6000, 7000, 8000, 9000, or about 10000 μm. Individualmetallic nanoparticles can have any suitable size. For example, alargest dimension of an individual metallic nanoparticle can be in arange of from about 25 nm to about 500 nm, about 50 nm to about 100 nm,or less than, equal to, or greater than about 25 nm, 30, 35, 40, 45, 50,55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130,135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195, 200,205, 210, 215, 220, 225, 230, 235, 240, 245, 250, 255, 260, 265, 270,275, 280, 285, 290, 295, 300, 305, 310, 315, 320, 325, 330, 335, 340,345, 350, 355, 360, 365, 370, 375, 380, 385, 390, 400, 405, 410, 415,420, 425, 430, 435, 440, 445, 450, 455, 460, 465, 470, 475, 480, 485,490, 495, or about 500 nm.

Etched fiber base 102 having metallic nanoparticle coating 104 disposedthereon can be deployed into an environment as a stand-alone component.Alternatively, etched fiber base 102 having metallic nanoparticle 104disposed thereon can be at least partially disposed within anothercomponent. For example, as shown in FIG. 1, etched fiber base 102 havingmetallic nanoparticle 104 disposed thereon is at least partiallycircumscribed and disposed in needle 106. In other embodiments, etchedfiber base 102 having metallic nanoparticle 104 disposed thereon can bedisposed at least partially within a container such as a straw.

FIGS. 2A-2B are schematic depictions of a needle 206 for a SERSsubstrate assembly (e.g., assembly 100 in FIG. 1). Needle 206 caninclude, for example, etched fiber base 202 and metallic nanoparticlecoating 204, which is disposed over at least a portion of the surface ofetched fiber base 202. Needle 206 at least partially circumscribesetched fiber base 202. Assembly 200 additionally includes in-situ filter208. Components 202 and 204, are similar to the corresponding componentsdescribed in reference to FIG. 1 and are connected in a similar fashion.

FIG. 2A shows needle 206 separate from a medium. FIG. 2B shows needle206 in a medium 210 with analyte 212. The medium in FIG. 2B can be agaseous medium, a liquid medium, a semi-solid medium, or a medium thatis a mixture of states. For example, the medium could be a gaseous orliquid solvent holding an analyte, such as a beverage, or could be theinterior of a fruit, vegetable, meat, a dairy product, a grain, or othersolid material being tested for analytes as discussed in depth below.

In-situ filter 208 caps needle 206, allowing the medium and analyte topass through in-situ filter 208 prior to reaching etched fiber base 202with metallic nanoparticle coating 204. In-situ filter 208 allowsanalyte to pass to etched fiber base 202 for analysis but preventsimpurities (i.e., other components of the medium or matrix) fromaffecting etched fiber base 202. Examples of impurities include apesticide, a metabolite, a pathogen, a bacteria, a fungi, a virus, anenzyme, a reactive oxygen species, and a mixture thereof. These arediscussed in more detail below. Filtration by in-situ filter 208 can bemechanical (e.g., filtration by size of particles or impurities due topore size of filter 208) or chemical (e.g., filter 208 could includeligands or other chemical components that are likely to capture certainimpurities). By reducing impurities, in-situ filter 208 also increasessensitivity of etched fiber base 202 to the analyte of interest.

Needle 206 can be a metallic (e.g., stainless steel copper, lead,chromium, tin, magnesium, aluminum, zinc, manganese, calcium, alloysthereof, and mixtures thereof), or plastic need suitable for puncturinga material for example, a hypodermic needle. Needle 206 can be a hollowneedle (e.g., an extraction needle shell). Needle 206 can have, forexample, a tapered end to allow for puncturing of a medium. In contrast,etched fiber base 202 can be blunt. In-situ filter 208 can be integralwith needle 206, and etched fiber base 202 can be inserted into needle206. In some embodiments, etched fiber base 202 is aligned with theoutside end of needle 206. Alternatively, etched fiber base 202 canextend beyond the outside end of needle 206 (see, for example, FIG. 10).

In-situ filter 208 can, for example, cover just an end of the needle206, or partially cover the sides of the needle 206, or can fill the endor a portion of needle 206, depending on the extent of protection ofetched fiber base 202 desired. In some embodiments, protection of etchedfiber base 202 with in-situ filter 208 allows for “naked” etched fiberbase 202 without metallic nanoparticle coating 204.

In-situ filter 208 can be a polymeric porous membrane attached to inneedle 206. In-situ filter 208 can be made, for example, by a polymercoating that can be cured to the end of needle 206. For example, in-situfilter 208 can be made of cellulose, nitrocellulose,polytetrafluoroethylene (PTFE), nylon, polycarbonate, acrylic basedpolymers, methacrylic based polymers, and combinations thereof. In-situfilter 208 can chemically or physically immobilize active absorbents orcatalytic compounds in the medium. Alternatively, the in-situ filter 208can remove or breakdown interfering compounds from the medium.

In-situ filter 208 can be a porous material so as to allow passage ofthe analyte through toward etched fiber base 202. This can also allowin-situ filter 208 to have a large surface area for absorbing orinteracting with impurities. FIG. 12 shows an SEM photograph of anexample in-situ filter. In-situ filter 208 can have, for example, anaverage pore size (across the longest point of the pore) of about 5 μmto about 35 μm (i.e., about 10 μm to about 25 μm, preferably from about12 μm to about 20 μm). In-situ filter 208 allows for passage of medium212 through into needle 206, where the medium is now filtered medium211, containing less impurities.

In-situ filter 208 can be pre-prepared and inserted into needle 206 orprepared directly in needle 206 via in-situ polymerization. The in-situfilter 208 can contain a cavity that allows insertion of the etchedfiber base 202.

In operation, surface enhanced Raman scattering substrate assembly 100or 200 can be used to detect an analyte. A method for using surfaceenhanced Raman scattering substrate assembly 100 or 200, illustrated inFIG. 3, can include contacting etched fiber base 102 or 202 havingmetallic nanoparticle 104 or 204 disposed thereon with a medium. Etchedfiber base 102 or 202 having metallic nanoparticle 104 or 204 disposedthereon can then be removed from contact with the medium and contactedwith a laser emission. A spectrum can then be generated and analyzed forthe presence or absence of an analyte. The amount of analyte present orthe concentration of the analyte in solution can also be quantitativelydetermined from the spectrum that is generated.

Surface enhanced Raman scattering substrate assembly 100 or 200 can beadapted to collect and subsequently detect an analyte that is in any oneof the gaseous phase, liquid phase, or solid phase. To that end, etchedfiber base 102 or 202 having metallic nanoparticle 104 or 204 disposedthereon can be disposed in a gaseous phase, a liquid phase, or a solidphase. In some embodiments, substrate assembly 100 or 200 can bedisposed in all three phases or any two of the three phasessimultaneously. For example, a first region of etched fiber base 102 or202 can be located in a gaseous phase, a second region of etched fiberbase 102 or 202 can be located in a liquid phase (e.g., in an organicliquid phase, in an aqueous liquid phase, or both), and a third regionof etched fiber base 102 or 202 can be located in a solid phase.According to some embodiments, each region of etched fiber base 102 or202 can be specifically configured for collection of analytes in aspecific phase. Disposing etched fiber base 102 or 202 across multiplephases can allow for simultaneous collection of analytes on etched fiberbase 102 or 202 across those phases.

After the analyte or analytes are collected on etched fiber base 102 or202, etched fiber base 102 or 202 can be removed for detection of theanalyte or analytes by SERS. Alternatively, SERS can be carried out insitu if, for example, substrate assembly 100 or 200 is configured toallow electromagnetic radiation to interact with etched fiber base 102or 202.

As an example, as shown in FIG. 1, etched fiber base 102 or 202 havingmetallic nanoparticle 104 or 204 disposed thereon is placed in a sealedenvironment and is disposed within a gaseous medium. Analytes ofinterest are collected on etched fiber base 102 or 202. Etched fiberbase 102 or 202 can then be analyzed via SERS either by removing etchedfiber base 102 or 202 from needle 106 or 206, or as mentioned above, insome cases can be analyzed through assembly 100 or 200 if the vial istransparent to electromagnetic radiation.

Surface enhanced Raman scattering substrate assembly 100 or 200 can beused in conjunction with many different types of analytes. Suitableexamples of mediums include a food, a beverage, a plant, an animal, or amixture thereof. Suitable examples of food include a vegetable, a fruit,a meat, a dairy product, a grain, and mixtures thereof. Suitableexamples of beverages include milk, beer, wine, water, juice, coffee,tea, and mixtures thereof. In still further embodiments, the medium canbe generated from a living organism. For example, the medium can beliving organism's breath. In some embodiments, the medium, and thereforethe analyte, can be heated to put the analyte into a gaseous phase fordetection.

The analyte can be any analyte of interest. For example, the analyte canbe a pesticide, a metabolite, a pathogen, a bacteria, a fungi, a virus,an enzyme, a reactive oxygen species, and a mixture thereof. Examples ofa suitable pesticide is O-Ethyl S-phenyl ethylphosphonodithioate(fonofos), thiabendazole, acetamiprid, (irontris(dimethyldithiocarbamate) (ferbam), phosmet, phorate, isocarbophos,and mixtures thereof. Examples of suitable metabolites include salicylicacid, phytoalexin, sulfonic acid, diphenyl sulfide, allyl methylsulfide, and a mixture thereof. In general, any compound includingsulfur may be an analyte of interest. Examples of a suitable enzymeinclude an enzyme including adenine (e.g., flavin adenine dinucleotide),nicotinamide adenine dinucleotide phosphate oxidase. Suitable bacteriafor examination may include a gram-positive bacteria, a gram-negativebacteria, and mixtures thereof. In some embodiments, the bacteria ischosen from Clostridium botulinum, Listeria monocytogenes, Acetic acidbacteria, Acidaminococcus, Acinetobacter baumannii, Agrobacteriumtumefaciens, Akkermansia muciniphila, Anaerobiospirillum, Anaerolineathermolimosa, Anaerolinea thermophila, Arcobacter, Arcobacter skirrowii,Armatimonas rosea, Azotobacter salinestris, Bacteroides, Bacteroidesfragilis, Bacteroides ureolyticus, Bacteroidetes, Bartonella japonica,Bartonella koehlerae, Bartonella taylorii, Bdellovibrio, Brachyspira,Bradyrhizobium japonicum, Caldilinea aerophile, Cardiobacterium hominis,Chaperone-Usher fimbriae, Christensenella, Chthonomonas calidirosea,Coxiella burnetiid, Cyanobacteria, Cytophaga, Dehalogenimonaslykanthroporepellens, Desulfurobacterium atlanticum, Devosia pacifica,Devosia psychrophila, Devosia soli, Devosia subaequoris, Devosiasubmarina, Devosia yakushimensis, Dialister, Dictyoglomus thermophilum,Enterobacter, Enterobacter cloacae, Enterobacter cowanii,Enterobacteriaceae, Enterobacteriales, Escherichia, Escherichia coli,Escherichia fergusonii, Escherichia hermannii, Fimbriimonasginsengisoli, Flavobacterium, Flavobacterium akiainvivens, Francisellanovicida, Fusobacterium necrophorum, Fusobacterium nucleatum,Fusobacterium polymorphum, Haemophilus felis, Haemophilus haemolyticus,Haemophilus influenzae, Haemophilus pittmaniae, Helicobacter, Kingellakingae, Klebsiella pneumoniae, Kluyvera ascorbate, Kluyveracryocrescens, Legionella, Legionella clemsonensis, Legionellapneumophila, Leptonema illini, Leptotrichia buccalis, Levilineasaccharolytica, Luteimonas aquatic, Luteimonas composti, Luteimonaslutimaris, Luteimonas marina, Luteimonas mephitis, Luteimonas vadose,Megamonas, Megasphaera, Meiothermus, Meiothermus timidus,Methylobacterium fujisawaense, Morax Axenfeld diplobacilli, Moraxella,Moraxella bovis, Moraxella osloensis, Morganella morganii, Mycoplasmaspumans, Neisseria cinereal, Neisseria gonorrhoeae, Neisseriameningitidis, Neisseria polysaccharea, Neisseria sicca, Nitrosomonaseutropha, Nitrosomonas halophila, Nonpathogenic organisms, OMPdb,Pectinatus, Pedobacter heparinus, Pelosinus, Propionispora,Proteobacteria, Proteus mirabilis, Proteus penneri, Pseudomonas,Pseudomonas aeruginosa, Pseudomonas luteola, Pseudoxanthomonasbroegbernensis, Pseudoxanthomonas japonensis, Rickettsia rickettsia,Salinibacter ruber, Salmonella, Salmonella bongori, Salmonella enterica,Samsonia, Selenomonadales, Serratia marcescens, Shigella, Shimwellia,Solobacterium moorei, Sorangium cellulosum, Sphaerotilus natans,Sphingomonas gei, Spirochaeta, Spirochaetaceae, Sporomusa,Stenotrophomonas, Stenotrophomonas nitritireducens, Thermotoganeapolitana, Thorselliaceae, Trimeric autotransporter adhesion,Vampirococcus, Verminephrobacter, Vibrio adaptatus, Vibrio azasii,Vibrio campbellii, Vibrio cholerae, Victivallis vadensis, Vitreoscilla,Wolbachia, Yersiniaceae, Zymophilus, strains thereof, and mixturesthereof.

Detection of any analyte can be used to assess many parameters of amedium. For example, the presence of pesticides can be used to determinewhether food is safe for consumption. Detection of certain analytes canbe used to generate a profile of a medium. For example, a medium such aswine can be analyzed for certain analytes that can be used to grade wineusing artificial analysis. This can be used to perform a uniformanalysis of the wine having the subjectivity of a human wine testerremoved. As a further example the medium can be a person's breath. Thebreath can be analyzed for certain analytes that may be indicators ofbad breath or certain health issues. Furthermore, the presence ofcertain analytes such as metabolites can be indicators of biotic stressin a plant. This can be monitored continuously to assess the health of aplant.

Surface enhanced Raman scattering substrate assembly 100 or 200 can bemanufactured in many suitable manners. For example, a fiber can beetched to form etched fiber base 102 or 202. As described herein, thefiber can be etched with a laser or by exposing the fiber to an etchant.The etchant can be an acid such as hydrochloric acid.

Following etching, the metallic nanoparticles are coated on the surfaceof etched fiber base 102 or 202. Coating can be accomplished by at leastpartially immersing etched fiber base 102 in a solution comprising themetal of the metallic nanoparticle. The metal in the solution is thenreduced thereon. In some embodiments in which metallic nanoparticlelayer 104 or 204 includes gold nanoparticles, the solution comprisingthe metal can be HAuCl₄.

In general, the assemblies 100, 200, disclosed herein allow for analysisof target compounds (analytes) in complex mediums such as food orbiological samples without complex sampling methods. An in-situmicro-extraction device used with a SERS needle can improve sensitivityto analytes with minimally invasive techniques.

EXAMPLES

Various embodiments of the present disclosure can be better understoodby reference to the following Examples which are offered by way ofillustration. The present disclosure is not limited to the Examplesgiven herein.

Example 1

In this Example, a highly sensitive surface enhanced Raman scatteringmethod coupled with headspace and solid phase micro-extraction (SPME) todetect volatile pesticide fonofos using an etched fiber having ametallic nanoparticle coating disposed thereon is described. Fonofos, orO-ethyl S-phenyl ethylphosphonodithiolate, is selected as a model fordetection using this method because of its volatility (i.e., boilingpoint is 130° C. at 0.1 mm Hg). It is one of the organophosphate soilinsecticides that can control pests such as corn rootworms. According toEnvironmental Protection Agency regulatory document, the oral exposureto fonofos can induce acethylcholinesterase inhibition and cause acutetoxicity. The chronic reference dose for fonofos is 0.002 mg/kg/day, thehealth reference level is 10 ppb, and the minimum reporting level is 0.5ppb. A gold-nanoparticle coated fiber was fabricated using a chemicaletching and coating method. The characterization of the fabricatedfibers and their performance in headspace-SPME and dip-SPME methodsfollowed by SERS analysis were determined in water and complex matrix(i.e. apple juice).

Materials. Analytical grade standard of fonofos (>99.9%), hydrogentetrachloroaurate hydrate (99.999%), and sodium chloride (>99.5%) wereprocured from Sigma Aldrich (St. Louis, Mo., USA). Hydrochloric acid(34%-37.5%), Acetonitrile (99.9%), ethanol (100%) and methanol (99.9%)were purchased from Fisher Scientific (Fair Lawn, N.J., USA). TheLangres® apple juice was purchased from local Stop & Shop supermarket(Amherst, Mass., USA). The stainless-steel wire (SUS304, φ140 μm) waspurchased from the Small Parts, Inc. Stock solution of fonofos wasprepared in acetonitrile at 100 ppm and further diluted by distilledwater or apple juice.

Preparation of gold nanoparticles-coated fibers. An acid etchingreaction was used to increase the roughness of a wire fiber as well asstrengthen the binding between the gold-nanoparticles coating and theporous stainless wire with the increased surface. The stainless-steelwire (5 cm) was washed with methanol, ethanol and distilled water in anultrasonic bath for 10 min respectively and then chemically etched inhydrochloric acid (37.5%) to create the roughness of the wire fiber. Theetched fiber was washed again with methanol and distilled water in theultrasonic bath for 5 min respectively, then dried at 60° C. The etchedfiber was then immersed into HAuCl₄ solution (0.05%, w/w) to introducegold to its porous surface as demonstrated in FIG. 3. The coatingreaction is the replacement reaction between iron and gold that is shownbelow.

Fe+[AuCl₄]⁻=Fe³⁺+Au+4Cl⁻

The surface morphology of acid-etched fiber and gold-nanoparticlescoated fiber were characterized under microscopes and SEM.

Detection of pesticides using headspace-SPME and dip-SPME methods. Eachtest pesticide stock solution of 100 mg/L (ppm) was prepared withacetonitrile and further diluted to needed concentrations (0.5 ppm to0.005 ppm) with distilled water or apple juice prior to use. 5 mL ofworking solution was mixed with 3 mL of 20% sodium chloride solution ina 16-mL vial with an open top polypropylene closure and PTFE/siliconesepta. The addition of 20% NaCl solution can increase the ionicstrengths and thus decreases the solubility of organic analytes in theaqueous phase in headspace-SPME detection. In the headspace-SPME method,the fiber was inserted through the silicon septum into the headspaceabove the working solution to extract the volatile compounds. Theextraction condition was 75° C. for 30 min. After extraction, the fiberwas fixed on a slide for SERS analysis. In the dip-SPME detection,working solution remains the same while the fiber dipped into theworking solution without salt for 30 min under room temperature. Thefiber was then air-dried and measured using Raman microscopy.

Instruments and data analysis. The surface morphology of etched fiberand gold-nanoparticles coated fiber were characterized by FEI Magellan400 scanning electron microscope (SEM, Hillsboro, Oreg.) with thevoltage of 5.0 kV.

A DXR Raman microscope (Thermo Fisher Scientific, Madison, Wis., U.S.A.)with a 780 nm laser and a 50× confocal microscope objective (0.8 mm spotdiameter and 2 cm⁻¹ spectral resolution) was used in this study. Eachspectrum was scanned from 2000 to 800 cm⁻¹ with 1 mW laser power and a50 mm slit width for 2 seconds integration time. OMNIC™ software version9.1 was used to control the Raman instrument. Fifteen scans wereselected from each fiber and then averaged by the software.

The Raman spectra were analyzed using Thermo Scientific TQ Analyst 8.0software. All Raman intensities were calculated from at least threereplicates and standard deviations were recorded. The peak at 1571 cm⁻¹Raman shift of fonofos was chosen for further characteristic analysisdue to its good consistency and least interference with the AuNPsbackground and apple juice signals.

Characterization of fiber substrate and fonofos SERS spectra. The etchedfiber has rough surface as shown in FIG. 4A to 4D. After replacementreaction, the coated fiber showed golden color which indicates thesuccessful coating of Au in FIG. 4E and FIG. 4F. Under SEM, thenanoparticles were at around 100 nm and evenly and densely distributedin FIG. 4G and FIG. 4H. This fabrication method shows great advantage asa simple and rapid way for coating nanoparticles onto a stainless-steelfiber comparing to other fabrication methods including laser ablation,annealing and chemical reaction layer by layer.

After the fiber was fabricated, its SERS-active capability andextraction efficiency were tested in 1 ppm fonofos water solution withdip and headspace methods. In headspace-SPME approach, 20% NaCl solutionwas added to the sample as the addition of salt usually increases theionic strengths and decreases the solubility of organic analytes in theaqueous phase. From FIG. 5, the fiber has minimal background noisebetween 800 to 2000 cm′ Raman shift, providing no interference topesticide signals. In dip and headspace tests, the four most obviouspeaks of fonofos on 1001, 1024, 1081 and 1576 cm⁻¹ Raman shift wereobserved and characterized in FIG. 5. The peak at 1576 cm⁻¹ isattributed to ν(C═C)phenyl stretch which is used for quantitativeanalysis later. The peaks at 1081, 1024 and 1001 cm′ are respectivelyattributed to ν(S—C phenyl)+δ(C—H)phenyl, δ(C—H)phenyl+ν(S—C phenyl),and δ(CCC)phenyl (20). Moreover, headspace method presents higherintensity of signals and minimal interference compared to dip method,indicating the advantage and feasibility of headspace approach forfonofos detection.

To investigate the sensitivity and quantitative reliability of themethod, the headspace-SPME-SERS were applied to detect fonofos ofvarious concentrations (0.005 ppm to 0.5 ppm) in water as shown FIG. 6A.The lowest detectable concentration at 5 ppb (0.005 ppm) was reached.Current SERS studies in detecting fonofos report higher detectableconcentration at 10 ppm, and their limit of detection ranges from 0.1ppm to 1 ppm. In comparison, the disclosed method offers a hugeimprovement on sensitivity due to the use of the headspace method forcapturing volatile fonofos. Peak intensity at 1576 cm⁻¹ was selected forquantitative analysis and the linear range was obtained from 0.025 ppmto 0.5 ppm in FIG. 6B. Fonofos concentration and Raman intensity presenta nice linear relation with coefficient of determination (R²) as 0.9883.The Limit of Detection (LOD) value was calculated to be 0.0052 ppmaccording to the equation of 3.3 σ/S, where σ is the standard deviationof the blank, and S is the slope of the calibration curve. The LOD valueis confirmed by the detection of 0.005 ppm (5 ppb) fonofos in FIG. 6A.The theoretical Limit of Quantification (LOQ) value can be extended to0.015 ppm according to the equation of 10 σ/S. Yet, the error barsrevealed that the method had large variations that needs to be furtherreduced. The variation may come from varied sizes and aggregations ofthe gold-nanoparticles on the fiber which may be improved by using astainless wire fiber with a higher quality and purity and furtheroptimizing the coating reaction conditions.

To further illustrate the advantage of headspace method in detecting avolatile pesticide in a real matrix, the headspace method was appliedand compared with the dip method to detect fonofos in apple juice. FromFIG. 7A, the dip-SPME-SERS method detected 50 ppb fonofos spiked inapple juice and cannot detect lower concentration at 10 ppb because itwas affected by the inferencing compounds from apple juice. On the otherhand, headspace-SPME-SERS detected 5 ppb fonofos spiked in apple juice(FIG. 7B). This data demonstrates the headspace method is more sensitiveand effective than the dip method when detecting fonofos in complexmatrices. It is because in the headspace, only volatile compounds occupythe space and have the chance to bind to the fiber. While in the dipdetection, other components from the sample matrix may bind to the fiberand cause interference. The lowest detectable concentration at 5 ppb ina food sample is comparable to the nano-liquid chromatography and thecommon GC method in complex samples detection, which are 5.3 ppb and 30ppb, respectively.

Example 2

SERS techniques were applied to monitor plant metabolites andphysiological changes to provide insight into biochemical plantresponses to biotic stress. The study was used to monitor SERS signalsof salicylic acid (SA), nicotinamide adenine dinucleotide phosphateoxidase (NADPH oxidase), and camalexin. Each molecule is SERS sensitiveand each SERS fingerprint is different based on their chemicalstructures. This allows for sensitive and selective monitoring ofresponse signals in plant matrixes without overlap in spectral analyses.The presence of NADPH oxidase is also associated with reactive oxidespecies (ROS) and SA and emerges rapidly within a host. NADPH oxidaseand SERS analyses allow for detection of plant responses to bioticstress within minutes. The presence of camalexin specifically indicatesa plant response to biotic stress and this relationship can be assessedusing SERS.

Tomato plant (Solanum lycopersicum) was fostered as a model. Fusariumproliferatum, a common tomato plant pathogen, was administered onto thesurface of the plant tissues. The plant response was monitored inreal-time in-situ locally and systemically with a field-based portableRaman instrument. SERS detection was performed by adding penetrable Goldnanoparticles (AuNPs) onto the surface of leaves or flowers. On fruits,a smart ‘SERS needle’ device was utilized which is composed of a realclinical needle and an inserted AuNPs coated fiber. SERS detection canbe carried out by removing the fiber from the plant tissue forfiber-based collections of SERS spectra. The SERS needle device alsoallows for the injection of biotic stress factors directly into thefruit using a syringe wherein it is possible to monitor biomolecularstress responses in real-time locally or systemically.

Example 3

To analyze a person's breath a straw was inserted into a vial, with thegold-nanoparticles coated fiber put inside the straw. This assembly isshown in FIG. 8. Three individuals were asked to blow in the straw for 1minute. Then the straw was folded to seal the breathing gas, and thefiber could be fully exposed to the gas. After 5 minutes of exposure andextraction, the fiber was measured with Raman scattering with 5 mW laserpower. Resulting spectra are shown in FIG. 9.

Example 4

To detect the presence of an analyte such as a pesticide in food asurface enhanced Raman scattering substrate assembly was placedpartially within a tomato. This is schematically shown in FIG. 10. Asshown in FIG. 10, the needle is positioned within a solid phase, liquidphase, and gaseous (e.g., headspace) phase of the tomato.

100 ppm of the pesticide thiabendazole was injected on one side of thetomato. The pesticide was left to translocate through the tomato for atime period of 48 hours. After 48 hours, the surface enhanced Ramanscattering substrate was used to detect the presence of the pesticide inthe tomato. Detection was performed across the solid, liquid, andgaseous phases although detection can be optionally limited to any onephase of sub-combination of phases. FIG. 11 is a spectrum showing thepresence of thiabendazole in the tomato.

Example 5

In Example 5, an in-situ filter was synthesized in a needle via in-situpolymerization using a high internal phase emulsion (HIPE) technique.First, dicyclopentadiene (DCPD) was pre-mixed with (poly(ethyleneglycol)-block-poly(propylene glycol)-block-poly(ethylene glycol(Pluronic1L-121) to create a mixture. Then, deionized water (DI) (16 mL)was added dropwise to the mixture under constant stirring to formhigh-internal phase emulsion. A solution of(H2IMes)(PCy3)Cl2Ru(3-phenyl-indenylid-1-ene) (M2) in toluene was addedto the solution as a catalyst.

The mixed emulsion was withdrawn into a needle shell and cured in-situat 80° C. The cured DCPD polymer formed a porous polymer network wasused as an in-situ filter. Prior to application, the cured HIPE filterwas washed in acetone to remove any un-cured monomers. The in-situfilter made in Example 5 can be seen in FIG. 12. Alternative monomericcompounds could be used for in-situ filter synthesis, such asdivinylbenzene (DVB), 2-ethylhexyl acrylate (EHA) and methacrylate(EHMA), butyl acrylate (BA) and isobornyl acrylate (IBA), methylmethacrylate (MMA), and vinylbenzyl chloride (VBC). An etched fibercould then be inserted into the needle with the DCPD filter for use inSERS analysis.

The terms and expressions that have been employed are used as terms ofdescription and not of limitation, and there is no intention in the useof such terms and expressions of excluding any equivalents of thefeatures shown and described or portions thereof, but it is recognizedthat various modifications are possible within the scope of theembodiments of the present disclosure. Thus, it should be understoodthat although the present disclosure has been specifically disclosed byspecific embodiments and optional features, modification and variationof the concepts herein disclosed may be resorted to by those of ordinaryskill in the art, and that such modifications and variations areconsidered to be within the scope of embodiments of the presentdisclosure.

ADDITIONAL EMBODIMENTS

The following exemplary embodiments are provided, the numbering of whichis not to be construed as designating levels of importance:

Embodiment 1 provides a surface enhanced Raman scattering substrateassembly for detecting an analyte, the assembly comprising:

an etched fiber base;

a metallic nanoparticle coating disposed over at least a portion of anexternal surface of the etched fiber base.

Embodiment 2 provides the assembly of Embodiment 1, wherein the etchedfiber base comprises stainless steel, copper, lead, chromium, tin,magnesium, aluminum, zinc, manganese, calcium, alloys thereof, andmixtures thereof.

Embodiment 3 provides the assembly of any one of Embodiments 1 or 2,wherein the etched fiber base is acid-etched.

Embodiment 4 provides the assembly of any one of Embodiments 1-3,wherein the metallic nanoparticle coating is disposed over about 50% toabout 100% of the total surface area of the etched fiber base.

Embodiment 5 provides the assembly of any one of Embodiments 1-4,wherein the metallic nanoparticle coating is dispersed over about 70% toabout 100% of the total surface area of the etched fiber base.

Embodiment 6 provides the assembly of any one of Embodiments 1-5,wherein the metallic nanoparticle coating is dispersed over about 90% toabout 98% of the total surface area of the etched fiber base.

Embodiment 7 provides the assembly of any one of Embodiments 1-6,wherein a surface area of the etched fiber base is greater than acorresponding fiber base that is free of etching.

Embodiment 8 provides the assembly of any one of Embodiments 1-7,wherein the etched fiber base has a length ranging from about 3 cm toabout 6 cm.

Embodiment 9 provides the assembly of any one of Embodiments 1-8,wherein the etched fiber base has a length ranging from about 4 cm toabout 5 cm.

Embodiment 10 provides the assembly of any one of Embodiments 1-9,wherein the etched fiber base has a width ranging from about 100 μm toabout 400 μm.

Embodiment 11 provides the assembly of any one of Embodiments 1-9,wherein the etched fiber base has a width ranging from about 0.5 cm toabout 5 cm.

Embodiment 12 provides the assembly of any one of Embodiments 1-11,wherein the etched fiber base is substantially cylindrically shaped.

Embodiment 13 provides the assembly of any one of Embodiments 1-12,wherein the metallic nanoparticle coating comprises a plurality ofmetallic nanoparticles.

Embodiment 14 provides the assembly of any one of Embodiments 1-13,wherein each of the plurality of metallic nanoparticles, independentlycomprise Ag₂O, elemental silver, elemental gold, elemental copper,elemental platinum, mixtures thereof, alloys thereof, or combinationsthereof.

Embodiment 15 provides the assembly of any one of Embodiments 1-14,wherein each of the plurality of metallic nanoparticles compriseselemental gold.

Embodiment 16 provides the assembly of any one of Embodiments 1-15,wherein at least one of the plurality of metallic nanoparticles is ananosphere, a nanochain, a nanoreef, a nanobox, or a nanostar.

Embodiment 17 provides the assembly of any one of Embodiments 1-16,wherein a largest dimension of at least one of the plurality of metallicnanoparticles has a largest dimension in a range of from about 25 nm toabout 500 nm.

Embodiment 18 provides the assembly of any one of Embodiments 1-17,wherein a largest dimension of at least one of the plurality of metallicnanoparticles has a largest dimension in a range of from about 50 nm toabout 100 nm.

Embodiment 19 provides the assembly of any one of Embodiments 1-18,further comprising a metallic microparticle coating dispersed over atleast a portion of the surface of the etched fiber base.

Embodiment 20 provides the assembly of any one of Embodiments 1-19,further comprising a needle circumscribing at least a portion of thefiber base.

Embodiment 21 provides the assembly of Embodiment 20, wherein the needlecomprises a metal.

Embodiment 22 provides the assembly of any one of Embodiments 1-21,wherein the assembly is further configured to detect the analyte in atleast one of a gaseous phase, a liquid phase and a solid phase.

Embodiment 23 provides the assembly of any one of Embodiments 1-22,wherein the analyte is chosen from a pesticide, a metabolite, apathogen, a bacteria, a fungi, a virus, an enzyme, a reactive oxygenspecies, and a mixture thereof.

Embodiment 24 provides the assembly of Embodiment 23, wherein thepesticide is chosen from O-Ethyl S-phenyl ethylphosphonodithioate,thiabendazole, acetamiprid, iron tris(dimethyldithiocarbamate), phosmet,phorate, isocarbophos, and mixtures thereof.

Embodiment 25 provides the assembly of Embodiment 23, wherein themetabolite is chosen from salicylic acid, phytoalexin, sulfonic acid,diphenyl sulfide, allyl methyl sulfide, and a mixture thereof.

Embodiment 26 provides the assembly of Embodiment 23, wherein the enzymeis chosen from flavin adenine dinucleotide, nicotinamide adeninedinucleotide phosphate oxidase, and mixtures thereof.

Embodiment 27 provides the assembly of Embodiment 23, wherein thebacteria is chosen from a gram-positive bacteria, a gram-negativebacteria, and mixtures thereof.

Embodiment 28 provides the assembly of Embodiment 27, wherein thebacteria is chosen from Clostridium botulinum, Listeria monocytogenes,Acetic acid bacteria, Acidaminococcus, Acinetobacter baumannii,Agrobacterium tumefaciens, Akkermansia muciniphila, Anaerobiospirillum,Anaerolinea thermolimosa, Anaerolinea thermophila, Arcobacter,Arcobacter skirrowii, Armatimonas rosea, Azotobacter salinestris,Bacteroides, Bacteroides fragilis, Bacteroides ureolyticus,Bacteroidetes, Bartonella japonica, Bartonella koehlerae, Bartonellataylorii, Bdellovibrio, Brachyspira, Bradyrhizobium japonicum,Caldilinea aerophile, Cardiobacterium hominis, Chaperone-Usher fimbriae,Christensenella, Chthonomonas calidirosea, Coxiella burnetiid,Cyanobacteria, Cytophaga, Dehalogenimonas lykanthroporepellens,Desulfurobacterium atlanticum, Devosia pacifica, Devosia psychrophila,Devosia soli, Devosia subaequoris, Devosia submarina, Devosiayakushimensis, Dialister, Dictyoglomus thermophilum, Enterobacter,Enterobacter cloacae, Enterobacter cowanii, Enterobacteriaceae,Enterobacteriales, Escherichia, Escherichia coli, Escherichiafergusonii, Escherichia hermannii, Fimbriimonas ginsengisoli,Flavobacterium, Flavobacterium akiainvivens, Francisella novicida,Fusobacterium necrophorum, Fusobacterium nucleatum, Fusobacteriumpolymorphum, Haemophilus felis, Haemophilus haemolyticus, Haemophilusinfluenzae, Haemophilus pittmaniae, Helicobacter, Kingella kingae,Klebsiella pneumoniae, Kluyvera ascorbate, Kluyvera cryocrescens,Legionella, Legionella clemsonensis, Legionella pneumophila, Leptonemaillini, Leptotrichia buccalis, Levilinea saccharolytica, Luteimonasaquatic, Luteimonas composti, Luteimonas lutimaris, Luteimonas marina,Luteimonas mephitis, Luteimonas vadose, Megamonas, Megasphaera,Meiothermus, Meiothermus timidus, Methylobacterium fujisawaense, MoraxAxenfeld diplobacilli, Moraxella, Moraxella bovis, Moraxella osloensis,Morganella morganii, Mycoplasma spumans, Neisseria cinereal, Neisseriagonorrhoeae, Neisseria meningitidis, Neisseria polysaccharea, Neisseriasicca, Nitrosomonas eutropha, Nitrosomonas halophila, Nonpathogenicorganisms, OMPdb, Pectinatus, Pedobacter heparinus, Pelosinus,Propionispora, Proteobacteria, Proteus mirabilis, Proteus penneri,Pseudomonas, Pseudomonas aeruginosa, Pseudomonas luteola,Pseudoxanthomonas broegbernensis, Pseudoxanthomonas japonensis,Rickettsia rickettsia, Salinibacter ruber, Salmonella, Salmonellabongori, Salmonella enterica, Samsonia, Selenomonadales, Serratiamarcescens, Shigella, Shimwellia, Solobacterium moorei, Sorangiumcellulosum, Sphaerotilus natans, Sphingomonas gei, Spirochaeta,Spirochaetaceae, Sporomusa, Stenotrophomonas, Stenotrophomonasnitritireducens, Thermotoga neapolitana, Thorselliaceae, Trimericautotransporter adhesion, Vampirococcus, Verminephrobacter, Vibrioadaptatus, Vibrio azasii, Vibrio campbellii, Vibrio cholerae,Victivallis vadensis, Vitreoscilla, Wolbachia, Yersiniaceae, Zymophilus,strains thereof, and mixtures thereof.

Embodiment 29 provides the assembly of any one of Embodiments 1-28,wherein the analyte is located in a medium.

Embodiment 30 provides the assembly of Embodiment 29, wherein the mediumcomprises a food, a beverage, a plant, an animal, or a mixture thereof.

Embodiment 31 provides the assembly of Embodiment 30, wherein the foodis chosen from a vegetable, a fruit, a meat, a dairy product, a grain,and mixtures thereof.

Embodiment 32 provides the assembly of Embodiment 30, wherein thebeverage is chosen from milk, beer, wine, water, juice, coffee, tea, andmixtures thereof.

Embodiment 33 provides the assembly of any one of Embodiments 1-32,further comprising:

a source of electromagnetic radiation in optical communication with themetallic nanoparticle coating; and

a detector for detecting a signal from the metallic nanoparticlecoating.

Embodiment 34 provides a method for detecting an analyte, the methodcomprising:

contacting the etched fiber base of any one of Embodiments 1-33 with themedium of any one of Embodiments 29-33;

contacting the medium with an electromagnetic emission;

detecting the analyte; and

generating a spectrum.

Embodiment 35 provides the method of Embodiment 34, further comprisingidentifying an analyte in the medium from the spectrum.

Embodiment 36 provides the method of Embodiment 35, further comprisingquantifying an amount of analyte present in the medium from thespectrum.

Embodiment 37 provides the method of any one of Embodiments 34-36,wherein the etched fiber base is located in at least one of a gaseousphase, a liquid phase, or a solid phase.

Embodiment 38 provides the method of any one of Embodiments 34-37,wherein the etched fiber base and the medium are located in a sealedenvironment.

Embodiment 39 provides the method of any one of Embodiments 34-38,wherein the electromagnetic emission is a laser emission.

Embodiment 40 provides the method of any one of Embodiments 34-39,wherein the analyte is an indicator of a response to biotic stress.

Embodiment 41 provides the method of any one of Embodiments 34-40,further comprising heating the analyte to a temperature sufficient toput the analyte into a gaseous phase.

Embodiment 42 provides a method of making the assembly of any one ofEmbodiments 1-41, the method comprising:

etching a fiber to form the etched fiber base; and

coating the metallic nanoparticles on the surface of the etched fiberbase.

Embodiment 43 provides the method of Embodiment 42, wherein the fiber isetched by exposing the fiber to an etchant.

Embodiment 44 provides the method of Embodiment 43, wherein the etchantis an acid.

Embodiment 45 provides the method of any one of Embodiments 43 or 44,wherein the acid is hydrochloric acid.

Embodiment 46 provides the method of any one of Embodiments 42-45,wherein coating the metallic nanoparticles on the surface of the etchedfiber base comprises at least partially immersing the etched fiber basein a solution comprising the metal of the metallic nanoparticle andreducing the metal in the solution.

Embodiment 47 provides the method of Embodiment 46, wherein the solutioncomprises HAuCl₄.

Embodiment 48 provides the assembly of Embodiment 1, further comprisinga needle circumscribing at least a portion of the fiber base, and anin-situ filter in the needle.

Embodiment 49 provides the assembly of Embodiment 1, wherein the in-situfilter comprises of cellulose, nitrocellulose, polytetrafluoroethylene(PTFE), nylon, polycarbonate, acrylic based polymers, methacrylic basedpolymers, and combinations thereof.

Embodiment 50 provides the assembly of Embodiment 1, wherein the in-situfilter comprises a plurality of pores each having a size of about 5 μmto about 35 μm.

Embodiment 51 provides the assembly of Embodiment 50, wherein thein-situ filter comprises a plurality of pores each having a size ofabout 10 μm to about 25 μm.

Embodiment 52 provides the assembly of Embodiment 1, wherein the in-situfilter is configured to filter impurities out of the medium.

Embodiment 53 provides the assembly of Embodiment 1, wherein the in-situfilter is configured to prevent impurities from reaching the fiber base.

Embodiment 54 provides the assembly of Embodiment 1, wherein the in-situfilter is located in an end of the needle.

Embodiment 55 provides the assembly of Embodiment 1, wherein the in-situfilter is located along the walls of the needle.

What is claimed is:
 1. A method for detecting an analyte, the method comprising: contacting an etched fiber base with a medium to collect the analyte from the medium on the etched fiber base, wherein a metallic nanoparticle coating is disposed over at least a portion of an external surface of the etched fiber base, the metallic nanoparticle coating comprising metallic nanoparticles that have a largest dimension of about 25 nm to about 500 nm; and contacting the etched fiber base comprising the analyte with an electromagnetic emission.
 2. The method of claim 2, further comprising identifying or quantifying an analyte in the medium from the spectrum.
 3. The method of claim 1, further comprising identifying an analyte in the medium from the spectrum.
 4. The method of claim 1, wherein the etched fiber base is located in at least one of a gaseous phase, a liquid phase, or a solid phase.
 5. The method of claim 1, wherein the etched fiber base and the medium are located in a sealed environment.
 6. The method of claim 1, wherein the electromagnetic emission is a laser emission.
 7. The method of claim 1, wherein the analyte is an indicator of a response to biotic stress.
 8. The method of claim 1, further comprising heating the analyte to a temperature sufficient to put the analyte into a gaseous phase.
 9. The method of claim 1, wherein the analyte is chosen from a pesticide, a metabolite, a pathogen, a bacteria, a fungi, a virus, an enzyme, a reactive oxygen species, and a mixture thereof.
 10. The method of claim 9, wherein the pesticide is chosen from O-Ethyl S-phenyl ethylphosphonodithioate, thiabendazole, acetamiprid, iron tris(dimethyldithiocarbamate), phosmet, phorate, isocarbophos, and mixtures thereof.
 11. The method of claim 9, wherein the metabolite is chosen from salicylic acid, phytoalexin, sulfonic acid, diphenyl sulfide, allyl methyl sulfide, and a mixture thereof.
 12. The method of claim 9, wherein the enzyme is chosen from flavin adenine dinucleotide, nicotinamide adenine dinucleotide phosphate oxidase, and mixtures thereof.
 13. The method of claim 9, wherein the bacteria is chosen from a gram-positive bacteria, a gram-negative bacteria, and mixtures thereof.
 14. The method of claim 13, wherein the bacteria is chosen from Clostridium botulinum, Listeria monocytogenes, Acetic acid bacteria, Acidaminococcus, Acinetobacter baumannii, Agrobacterium tumefaciens, Akkermansia muciniphila, Anaerobiospirillum, Anaerolinea thermolimosa, Anaerolinea thermophila, Arcobacter, Arcobacter skirrowii, Armatimonas rosea, Azotobacter salinestris, Bacteroides, Bacteroides fragilis, Bacteroides ureolyticus, Bacteroidetes, Bartonella japonica, Bartonella koehlerae, Bartonella taylorii, Bdellovibrio, Brachyspira, Bradyrhizobium japonicum, Caldilinea aerophile, Cardiobacterium hominis, Chaperone-Usher fimbriae, Christensenella, Chthonomonas calidirosea, Coxiella burnetiid, Cyanobacteria, Cytophaga, Dehalogenimonas lykanthroporepellens, Desulfurobacterium atlanticum, Devosia pacifica, Devosia psychrophila, Devosia soli, Devosia subaequoris, Devosia submarina, Devosia yakushimensis, Dialister, Dictyoglomus thermophilum, Enterobacter, Enterobacter cloacae, Enterobacter cowanii, Enterobacteriaceae, Enterobacteriales, Escherichia, Escherichia coli, Escherichia fergusonii, Escherichia hermannii, Fimbriimonas ginsengisoli, Flavobacterium, Flavobacterium akiainvivens, Francisella novicida, Fusobacterium necrophorum, Fusobacterium nucleatum, Fusobacterium polymorphum, Haemophilus felis, Haemophilus haemolyticus, Haemophilus influenzae, Haemophilus pittmaniae, Helicobacter, Kingella kingae, Klebsiella pneumoniae, Kluyvera ascorbate, Kluyvera cryocrescens, Legionella, Legionella clemsonensis, Legionella pneumophila, Leptonema illini, Leptotrichia buccalis, Levilinea saccharolytica, Luteimonas aquatic, Luteimonas composti, Luteimonas lutimaris, Luteimonas marina, Luteimonas mephitis, Luteimonas vadose, Megamonas, Megasphaera, Meiothermus, Meiothermus timidus, Methylobacterium fujisawaense, Morax-Axenfeld diplobacilli, Moraxella, Moraxella bovis, Moraxella osloensis, Morganella morganii, Mycoplasma spumans, Neisseria cinereal, Neisseria gonorrhoeae, Neisseria meningitidis, Neisseria polysaccharea, Neisseria sicca, Nitrosomonas eutropha, Nitrosomonas halophila, Nonpathogenic organisms, OMPdb, Pectinatus, Pedobacter heparinus, Pelosinus, Propionispora, Proteobacteria, Proteus mirabilis, Proteus penneri, Pseudomonas, Pseudomonas aeruginosa, Pseudomonas luteola, Pseudoxanthomonas broegbernensis, Pseudoxanthomonas japonensis, Rickettsia rickettsia, Salinibacter ruber, Salmonella, Salmonella bongori, Salmonella enterica, Samsonia, Selenomonadales, Serratia marcescens, Shigella, Shimwellia, Solobacterium moorei, Sorangium cellulosum, Sphaerotilus natans, Sphingomonas gei, Spirochaeta, Spirochaetaceae, Sporomusa, Stenotrophomonas, Stenotrophomonas nitritireducens, Thermotoga neapolitana, Thorselliaceae, Trimeric autotransporter adhesion, Vampirococcus, Verminephrobacter, Vibrio adaptatus, Vibrio azasii, Vibrio campbellii, Vibrio cholerae, Victivallis vadensis, Vitreoscilla, Wolbachia, Yersiniaceae, Zymophilus, strains thereof, and mixtures thereof.
 15. The method of claim 1, wherein the medium comprises a food, a beverage, a plant, an animal, a breath, or a mixture thereof.
 16. The method of claim 15, wherein the food is chosen from a vegetable, a fruit, a meat, a dairy product, a grain, and mixtures thereof.
 17. The method of claim 15, wherein the beverage is chosen from milk, beer, wine, water, juice, coffee, tea, and mixtures thereof.
 18. The method of claim 1, further comprising detecting the analyte.
 19. The method of claim 1, further comprising generating a spectrum.
 20. The method of claim 1, wherein the etched fiber base has a length range from about 3 cm to about 6 cm. 